Overexpression of hSav1 promotes Mst1-induced apoptosis in HeLa cells / 中华肿瘤杂志
Zhonghua zhong liu za zhi
; (12): 481-484, 2009.
Article
in Zh
| WPRIM
| ID: wpr-293084
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To elucidate the effect of hSav1 expression on Mst1-mediated apoptosis in HeLa cells.</p><p><b>METHODS</b>Plasmids pCMV-HA-hSav1 and pcDNA/4TO-Flag-Mst1 were constructed and cotransfected into HeLa cells. Triple immunofluorescent labeling of hSav1, Mst1 and nucleus was performed to determine their subcellular localization. Plasmids pCMV-HA-hSav1 and/or pcDNA/4TO-Flag-Mst1 were transfected into HeLa cells, and 36 hours later cisplatin (50 micromol/L) as a pro-apoptotic agent was added for 14 hours. Cell apoptosis was analyzed by annexin V/PI assay.</p><p><b>RESULTS</b>Plasmids pCMV-HA-hSav1 and pcDNA/4TO-Flag-Mst1 were constructed and the authenticity of constructs was verified by sequencing. The binding in vitro showed that hSav1 could be detect from the anti-Mst1 immunoprecipitation complex. The immunofluorescent labeling showed that hSav1 and Mst1 had the same localization in cells. Overexpressed protein hSav1 did not induce a significant cell apoptosis. However, co-expression of hSav1 with Mst1 resulted in a significant increase of apoptosis above the level seen with Mst1 alone (24.5% +/- 2.4% vs. 39.3% +/- 4.0%, P < 0.05).</p><p><b>CONCLUSION</b>Our findings indicate that hSav1 is a newly identified protein that interacts with Mst1 and augments Mst1-mediated apoptosis.</p>
Full text:
1
Index:
WPRIM
Main subject:
Plasmids
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HeLa Cells
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Transfection
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Proto-Oncogene Proteins
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Hepatocyte Growth Factor
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Apoptosis
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Cell Cycle Proteins
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Cytoplasm
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Genetics
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Metabolism
Type of study:
Prognostic_studies
Limits:
Humans
Language:
Zh
Journal:
Zhonghua zhong liu za zhi
Year:
2009
Type:
Article